Noda Takahiro, Kikuchi Motoshi, Kaidzu Sachiko, Yashiro Takashi
Division of Histology and Cell Biology, Department of Anatomy, Jichi Medical School, Tochigi, Japan.
Anat Rec A Discov Mol Cell Evol Biol. 2003 Jun;272(2):548-55. doi: 10.1002/ar.a.10065.
Hormone-producing cells in the rat anterior pituitary gland are not randomly distributed; rather, there are specific topographic affinities among five cell types (Noda et al., Acta Histochem. Cytochem. 2001;34:313-319). In this study we reconstructed these affinities, at least partially, in primary monolayer culture. Pituitary cells collected from adult male rats were enzymatically dispersed and cultured for 72 hr at a density of 1 x 10(5) cells/cm(2). We double-immunostained cells using antibodies against hormones, and then used confocal laser microscopy to examine the ability of the cells to attach to each other. We also statistically analyzed the affinity of all combinations of the five types of hormone-producing cells. We observed clusters by electron microscopy to identify junctional complexes between the cells. Confocal laser microscopy indicated that the features and attachment patterns of hormone-producing cells in vivo were similar to those in vitro. Statistical analyses revealed that the rates at which the five types of hormone-producing cells attached to growth hormone (GH)-, prolactin (PRL), and luteinizing hormone (LH)-producing cells were unequal, which suggests there are specific topographic affinities. The specific rates of adrenocorticotropic hormone (ACTH)-producing cell attachment to GH cells, LH to PRL cells, and PRL to LH cells were high, whereas that of PRL attachment to PRL cells was low. In addition, the rates correlated with the data from our previous in vivo study. Ultrastructural observations revealed few junctional complexes between hormone-producing cells. These results indicate that anterior pituitary hormone-producing cells can attach to specific types of cells by means of specific and/or nonspecific adhesion factors, and can reconstruct the topographic nature of the pituitary gland.
大鼠垂体前叶中产生激素的细胞并非随机分布;相反,五种细胞类型之间存在特定的拓扑亲和性(野田等人,《组织化学与细胞化学学报》,2001年;34:313 - 319)。在本研究中,我们在原代单层培养中至少部分地重建了这些亲和性。从成年雄性大鼠收集的垂体细胞经酶分散后,以1×10⁵个细胞/cm²的密度培养72小时。我们使用针对激素的抗体对细胞进行双重免疫染色,然后利用共聚焦激光显微镜检查细胞彼此附着的能力。我们还对五种产生激素的细胞的所有组合的亲和性进行了统计分析。我们通过电子显微镜观察细胞簇以识别细胞间的连接复合体。共聚焦激光显微镜显示,体内产生激素的细胞的特征和附着模式与体外相似。统计分析表明,五种产生激素的细胞附着于生长激素(GH)、催乳素(PRL)和促黄体生成素(LH)产生细胞的速率不相等,这表明存在特定的拓扑亲和性。促肾上腺皮质激素(ACTH)产生细胞附着于GH细胞、LH附着于PRL细胞以及PRL附着于LH细胞的特定速率较高,而PRL附着于PRL细胞的速率较低。此外,这些速率与我们之前体内研究的数据相关。超微结构观察显示,产生激素的细胞之间很少有连接复合体。这些结果表明,垂体前叶产生激素的细胞可通过特定和/或非特定的黏附因子附着于特定类型的细胞,并可重建垂体的拓扑性质。